The present invention relates to an anthelmintic composition comprising a mixture of a macrolide antibiotic selected from those of the B-41, C-076 and 22,23-dihydro C-076 series (all of which are known to have anthelmintic activity) with certain other known anthelmintic agents, whereby the anthelmintic activity of the composition is synergistically enhanced.
The compounds referred to herein as "B-41 series antibiotics," "C-076 series antibiotics" and "22,23-dihydro C-076 series antibiotics" are a group of macrolide antibiotics which, despite their different nomenclatures (arising from their different methods of production by various microorganisms), have very closely related molecular structures and activities.
The B-41 series antibiotics were originally isolated from a culture broth of Streptomyces B-41-146 strain (deposited with the Fermentation Research Institute, Agency of Industrial Science and Technology, Ministry of International Trade and Industry, Japan, whence it is available under the Accession No. 1438).
Since the original discovery of the B-41 series compounds, described in British patent specification No. 1,390,336, wherein nine compounds were characterized, a number of other compounds from the same series have been isolated from a culture broth of the same microorganism. As disclosed in this British patent specification, these compounds may be prepared by cultivating a microorganism of the genus Streptomyces, preferably Streptomyces B-41-146 strain, in a suitable culture medium for a period of from 5 to 10 days at about 28.degree. C. under aerobic conditions, after which the culture broth is filtered through diatomaceous earth, the cake obtained is extracted with methanol and then with hexane to given an oily substance and finally the substance is fractionated by column chromatography through silica gel.
The B-41 series compounds thus include compounds having the following formulae (I) or (II), in which the groups represented by R.sup.1 -R.sup.6 are as defined in Tables 1 and 2. ##STR1##
TABLE 1 ______________________________________ B-41 R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5 ______________________________________ .alpha..sub.1 (A.sub.3) H H CH.sub.3 CH.sub.3 OH .alpha..sub.2 (B.sub.2) H H CH.sub.3 CH.sub.3 OCH.sub.3 .alpha..sub.3 (A.sub.4) H H C.sub.2 H.sub.5 CH.sub.3 OH .alpha..sub.4 (B.sub.3) H H C.sub.2 H.sub.5 CH.sub.3 OCH.sub.3 (D) H H i-C.sub.3 H.sub.7 CH.sub.3 OH (G) H H i-C.sub.3 H.sub.7 CH.sub.3 OCH.sub.3 .alpha..sub.5 (A.sub.2) OH MH CH.sub.3 CH.sub.3 OH .alpha..sub.6 (B.sub.1) OH MH CH.sub.3 CH.sub.3 OCH.sub.3 .alpha..sub.7 OH MH C.sub.2 H.sub.5 CH.sub.3 OH .alpha..sub.8 OH MH C.sub.2 H.sub.5 CH.sub.3 OCH.sub.3 .alpha..sub.9 (C.sub.1) H H CH.sub.3 PC OH .alpha..sub.10 (C.sub.2) H H C.sub.2 H.sub.5 PC OH (F) H H i-C.sub.3 H.sub.7 PC OH ______________________________________
In this Table, the following abbreviations are used:
iC.sub.3 H.sub.7 --means an isopropyl group; PA1 MH--means a 2-methylhexanoyloxy group of formula ##STR2## and PC--means a 2-pyrrolylcarbonyloxymethyl group of formula ##STR3## PA1 1.52 (singlet, 14--CH.sub.3); PA1 1.86 (broad singlet, 4--CH.sub.3); PA1 3.94 (doublet, J=6.2 Hz, 6--H); PA1 4.63 (singlet, 26--CH.sub.2); PA1 4.91 (broad triplet, J=8 Hz). PA1 1.59 (singlet, 14--CH.sub.3); PA1 1.81 (broad singlet, 4--CH.sub.3); PA1 3.06 (singlet, 5--OCH.sub.3); PA1 4.12 (doubled doublet, J=4.5 and 12 Hz, 26--CH.sub.2); PA1 4.30 (doubled doublet, J=6 and 12 Hz, 26--CH.sub.2); PA1 4.87 (broad triplet, J=7 Hz, 15--CH.dbd.); PA1 6.23 (doubled doublet, J=11 and 12 Hz, 10--CH.dbd.); PA1 6.43 (doublet, J=11 Hz, 9--CH.dbd.). PA1 1.48 (singlet, 4--CH.sub.3); PA1 3.91 (doublet, J=6 Hz, 6--H); PA1 4.60 (singlet, 26--CH.sub.2); PA1 6.1-6.3 (1H, multiplet); PA1 6.8-7.0 (2H, multiplet). PA1 1.50 (singlet, 14--CH.sub.3); PA1 1.79 (broad singlet, 4--CH.sub.3); PA1 3.44 (singlet, 5--OCH.sub.3); PA1 4.59 (singlet, 26--CH.sub.2); PA1 4.89 (broad triplet, J=8 Hz, 15--H). PA1 (a) one or more anthelmintic agents selected from the group consisting of the B-41 series antibiotics, the C-076 series antibiotics and the 22,23-dihydro C-076 series antibiotics; and PA1 (b) one or more other anthelmintic agents selected from the group consisting of benzimidazole, salicylamide and isoquinoline compounds. PA1 (b) one or more other anthelmintic agents selected from the group consisting of benzimidazole, salicylamide and isoquinoline compounds. PA1 2-(Methoxycarbonylamino)benzimidazole; PA1 5-Butyl-2-(methoxycarbonylamino)benzimidazole; PA1 5-Propoxy-2-(methoxycarbonylamino)benzimidazole; PA1 5-Ethoxy-2-(ethoxycarbonylamino)benzimidazole; PA1 5-Propylthio-2-(methoxycarbonylamino)benzimidazole; PA1 5-Phenylthio-2-(methoxycarbonylamino)benzimidazole; PA1 5-Phenylsulphinyl-2-(methoxycarbonylamino)benzimidazole; PA1 5-(2,4-Dichlorophenoxy)-6-chloro-2-methylthiobenzimidazole; PA1 6-Chloro-5-(2,3-dichlorophenoxy)-2-methylthiobenzimidazole; PA1 2-(4-Thiazolyl)benzimidazole; and PA1 5-Isopropoxycarbonylamino-2-(4-thiazolyl)benzimidazole. PA1 5-Chloro-N-(2-chloro-4-nitrophenyl)salicylamide; PA1 3,5-Diiodo-N-(3-chloro-4-p-chlorophenoxyphenyl)-salicylamide; PA1 3,5-Diiodo-N-[5-chloro-2-methyl-4-(.alpha.-cyano-4-chlorobenzyl)phenyl]sali cylamide; PA1 3,5,6-Trichloro-N-(3,5-dichloro-2-hydroxyphenyl)-salicylamide; PA1 2-Acetoxy-3,5-diiodo-N-(p-chlorophenyl)benzamide; and PA1 2-Acetoxy-3-bromo-5-chloro-N-(p-bromophenyl)-thiobenzamide. PA1 2-Cyclohexylcarbonyl-4-oxo-1,2,3,6,7,11b-hexahydro-4H-pyrazino[2,1-a]isoqui noline; and PA1 2-Benzoyl-4-oxo-1,2,3,6,7,11b-hexahydro-4H-pyrazino[2,1-a]isoquinoline. PA1 Haemonchus; PA1 Trichostrongylus; PA1 Ostertagia; PA1 Nematodirus; PA1 Cooperia; PA1 Ascaris; PA1 Bunostomum; PA1 Oesophagostomum; PA1 Chabertia; PA1 Trichuris; PA1 Strongylus; PA1 Trichonema; PA1 Dictyocaulus; PA1 Capillaria; PA1 Heterakis; PA1 Toxocara; PA1 Ascaridia; PA1 Oxyuris; PA1 Ancylostoma; PA1 Uncinaria; PA1 Toxascaris; and PA1 Parascaris. PA1 Taenia saginata; PA1 Hymenolepis diminuta; PA1 Hymenolepis nana; PA1 Moniezia benedeni; PA1 Diphyllobothrium latum; PA1 Diphyllobothrium erinacei; PA1 Echinococcus glanulosus; and PA1 Echinococcus multilocularis, PA1 Fasciola hepatica; PA1 Fasciola gigantica; PA1 Paragonimus westermanii; PA1 Fasciolopsis buski; PA1 Eurytrema pancreaticum; PA1 Eurytrema coelomaticum; PA1 Clonorchis sinensis; PA1 Schistosoma japonicum; PA1 Schistosoma haematobium; and PA1 Schistosoma mansoni.
TABLE 2 ______________________________________ B-41 R.sup.3 R.sup.6 ______________________________________ .beta..sub.1 (A.sub.1) CH.sub.3 CH.sub.2 OH .beta..sub.2 (A.sub.5) C.sub.2 H.sub.5 CH.sub.2 OH (E) i-C.sub.3 H.sub.7 CH.sub.2 OH ______________________________________
Of the compounds shown above, those identified as A.sub.1, A.sub.2, A.sub.3, A.sub.4, A.sub.5, B.sub.1, B.sub.2, B.sub.3, C.sub.1 and C.sub.2 are described in British patent specification No. 1,390,336. Those compounds identified by .alpha. or .beta. are described in The Journal of Antibiotics, 29(3), 76-14 to 76-16 and 29(6), 76-35 to 76-42 and the use of these compounds as anthelmintic agents is described in European Patent Publication No. 2916. Compound B-41D is described in British patent specification No. 2,056,986.
Compounds B-41E and B-41F are described in Japanese Patent Application No. 153,141/80 and Compound B-41G is described in Japanese patent application No. 7091/81.
All of these compounds were obtained from a culture broth of the microorganism Streptomyces strain B-41-146 in the form of an amorphous powder. The properties of Compounds B-41E, B-41F and B-41G are given below.